Bush Susan M, Krysan Patrick J
Genome Center of Wisconsin and Department of Horticulture, 1575 Linden Drive, University of Wisconsin, Madison, WI 53706, USA.
J Exp Bot. 2007;58(8):2181-91. doi: 10.1093/jxb/erm092. Epub 2007 May 22.
Loss-of-function, dominant-negative, and change-of-function genetic approaches were used to investigate the role played by the Arabidopsis mitogen-activated protein (MAP) kinase MPK6 throughout development. Plants homozygous for T-DNA null alleles of MPK6 displayed reduced male fertility and abnormal anther development. In addition, a portion of the seed produced by mpk6 plants was found to contain embryos that had burst out of their seed coats. To address potential functional redundancy, a dominant-negative version of MPK6 was constructed by changing the TEY activation loop motif to the amino acid sequence AEF. Plants expressing MPK6AEF via the MPK6 native promoter were found to produce excessive stomata, consistent with the recently described role of MPK6 in stomatal patterning. A novel floral phenotype characterized by abnormal sepal development was also observed in MPK6AEF lines. The gene expression pattern of the MPK6 native promoter was determined using a YFP-MPK6 fusion construct, and expression was observed throughout most plant tissues, consistent with a role for MPK6 in multiple developmental processes. The YFP-MPK6 construct was found to rescue the fertility phenotype of mpk6 null alleles, indicating that the fusion protein retains its biological activity. It was also observed, however, that plants expressing YFP-MPK6 displayed reduced apical dominance and a shortening of inflorescence internodes. These results suggest that the YFP tag modifies the activity of MPK6 in a manner that affects inflorescence development but not anther development. Taken together, the present results indicate that MPK6 is involved in the regulation of multiple aspects of plant development.
采用功能缺失、显性负性和功能改变的遗传方法来研究拟南芥促分裂原活化蛋白(MAP)激酶MPK6在整个发育过程中所起的作用。MPK6的T-DNA无效等位基因纯合的植株表现出雄性育性降低和花药发育异常。此外,发现mpk6植株产生的一部分种子含有已冲破种皮的胚。为了解决潜在的功能冗余问题,通过将TEY激活环基序改变为氨基酸序列AEF构建了MPK6的显性负性版本。通过MPK6天然启动子表达MPK6AEF的植株被发现产生过多的气孔,这与最近描述的MPK6在气孔模式形成中的作用一致。在MPK6AEF株系中还观察到一种以萼片发育异常为特征的新型花表型。使用YFP-MPK6融合构建体确定了MPK6天然启动子的基因表达模式,并且在大多数植物组织中都观察到了表达,这与MPK6在多个发育过程中的作用一致。发现YFP-MPK6构建体挽救了mpk6无效等位基因的育性表型,表明融合蛋白保留了其生物学活性。然而,也观察到表达YFP-MPK6的植株表现出顶端优势降低和花序节间缩短。这些结果表明,YFP标签以影响花序发育但不影响花药发育方式改变了MPK6的活性。综上所述,目前的结果表明MPK6参与了植物发育多个方面的调控。